Weekly dosing regimens for anti-cd30 vc-pab-mmae antibody drug-conjugates

ABSTRACT

Methods for the treatment of CD30-expressing cancers are provided. The methods comprise administering to a subject in need thereof a weekly dose of from about 0.8 mg/kg to about 1.8 mg/kg of an antibody-drug conjugate compound having formula (I); or a pharmaceutically acceptable salt thereof; wherein: mAb is an anti-CD30 antibody unit, S is a sulfur atom of the antibody, A- is a Stretcher unit, and p is from about 3 to about 5.

This application claims the benefit of U.S. Provisional App. No.61/143,713 filed Jan. 9, 2009; U.S. Provisional App. No. 61/152,205filed Feb. 12, 2009; U.S. Provisional App. No. 61/175,719 filed May 5,2009 and U.S. Provisional App. No. 61/264,222 filed Nov. 24, 2009, eachof which is incorporated by reference in its entirety for all purposes.

REFERENCE TO A SEQUENCE LISTING

This application includes a sequence listing submitted herewith as atext file named “Seqlist003000605PC.txt” created on Jan. 6, 2010, andcontaining 20,480 bytes. The material contained in this text file isincorporated by reference in its entirety for all purposes.

FIELD

The present invention relates, inter alia, to methods for the treatmentof a CD30-expressing hematologic cancer comprising administering to asubject in need thereof a weekly dose of from about 0.8 mg/kg to about1.8 mg/kg of an anti-CD30 vc-PAB MMAE antibody-drug conjugate.

BACKGROUND

Hodgkin lymphoma (HL) is a neoplasm of lymphoid tissue that is definedhistopathologically by the presence of the malignantHodgkin-Reed-Sternberg (HRS) cells. The characteristic surface antigenexpressed on HRS cells is CD30. There are an estimated 8,000 new HLcases diagnosed annually in the United States and Canada. Advances inthe use of combined chemotherapy and radiotherapy in HL over the pasthalf-century have resulted in a durable remission rate of approximately70%. However, these multi-agent regimens confer a significant morbidityon patients, including secondary malignancies, cardiac disease, andinfertility. Furthermore, approximately 30% of patients presenting withHL will become refractory to initial therapy or will relapse. Salvagechemotherapy regimens and autologous stem cell transplant (ASCT) aresecondary options for these patients, but both are associated withsignificant morbidity and limited long term disease control. Patientswho relapse after ASCT or are ineligible for salvage therapy have a verypoor prognosis. Currently, there is a lack of well-tolerated,efficacious treatment options for these patients. Thus, there continuesto be an unmet medical need for patients suffering from HL and otherCD30 expressing cancers. The present invention addresses this and otherneeds.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: Exemplary administration schedule for an antibody drugconjugate. The antibody drug conjugate is administered in a singleweekly dose, three out of 4 weeks, every 28 days.

FIG. 2: Exemplary administration schedule for an antibody drugconjugate. The antibody drug conjugate is administered in a singleweekly dose, three out of 4 weeks. Following an evaluation indicatingthat the subject has no detectable signs of cancer, the administrationschedule is reduced to once every three to six weeks.

DETAILED DESCRIPTION OF EXEMPLARY EMBODIMENTS A. General Introduction

The present invention provides, inter alia, methods for treating aCD30-expressing hematologic cancer. The present inventors havediscovered that a weekly therapeutic regimen with the anti-CD30antibody-drug conjugate cAC10-(MC-vc-PAB-MMAE)₄ at doses of about 0.8mg/kg to about 1.2 mg/kg provides a surprisingly efficacious therapeuticregimen, e.g., it allows for multiple objective responses within a shorttime period and has an acceptable tolerability profile despite frequentdosing. That such a substantial dose of the antibody-drug conjugate canbe provided at such frequent intervals and result in a high responserate and acceptable toxicity profile is a surprising finding. In a phaseI study, multiple complete responses were observed within 8 weeks oftreatment. The weekly dosing regimen has been observed to provide a highrate of response in a short time period. The high rate of response isalso suprising for a drug delivered, not as a combination therapy, butas a monotherapy, for the treatment of cancer. Accordingly, the presentmethods provide, inter alia, a weekly dosing regimen for administeringan anti-CD30 vc-PAB-MMAE antibody-drug conjugate to a subject. In someembodiments, the weekly dosing regimen increases the subject'sprobability of responding to the therapy as compared to other dosingregimens (e.g., as compared to an every three week dosing regimen). Insome embodiments, the dosing regimen does not increase the subject'sprobability of suffering from an adverse event (including a doselimiting toxicity) as compared to other dosing regimens (e.g., ascompared to an every three week dosing regimen) despite the fact thatthe total dose of antibody drug conjugate over a treatment cycle may begreater for the weekly dosing regimen as compared to other dosingregimens. The present invention also provides, inter alia, maintenancetherapy following the weekly dosing regimen.

For clarity of disclosure, and not by way of limitation, the detaileddescription of the invention is divided into the subsections whichfollow.

B. Summary

The present invention is based, inter alia, on the discovery that weeklydosing of a cAC10-MC-vc-PAB-MMAE antibody drug conjugate (Sanderson etal., Clinical Cancer Research 2005 11:843-852, incorporated by referenceherein in its entirety and for all purposes) provides a surprisinglyhigh rate of response with an acceptable tolerability profile.

C. Definitions and Abbreviations

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art pertinent to the methods and compositions described. As usedherein, the following terms and phrases have the meanings ascribed tothem unless specified otherwise.

The term “inhibit” or “inhibition of” as used herein means to a reduceby a measurable amount, or to prevent entirely.

The transitional phrase “consisting essentially of” as used hereinlimits the scope of a claim to the specified active agents or steps andthose additional active agents and steps that do not materially affectthe properties of the specificed active agents.

The term “agent” as used herein means an element, compound, or molecularentity, including, e.g., a pharmaceutical, therapeutic, or pharmacologiccompound. Agents can be natural or synthetic or a combination thereof.

An “anti-cancer agent” is an agent that exerts a cytotoxic or cytostaticeffect on cancer cells either alone or in combination with another agentas part of a treatment regimen. An “anti-cancer agent” can slow, stop,or reverse the progression of cancer in a subject. For example, ananti-cancer agent is an agent that can inhibit tumor growth, arresttumor growth, and/or cause the regression of already existing tumors.Anti-inflammatory agents or other agents administered to a subject withcancer to treat symptoms associated with cancer, but not the underlyingcancer itself, including, for example inflammation, pain, weight loss,and general malaise are not considered anti-cancer agents.

“Cytotoxic effect,” in reference to the effect of an agent on a cell,means killing of the cell. “Cytostatic effect” means an inhibition ofcell proliferation. A “cytotoxic agent” means an agent that has acytotoxic or cytostatic effect on a cell, thereby depleting orinhibiting the growth of, respectively, cells within a cell population.

The term “deplete,” in the context of the effect of an anti-CD30antibody-drug conjugate on CD30-expressing cells, refers to a reductionor elimination of the CD30-expressing cells.

As used herein, the terms “treatment” or “treat” refer to slowing,stopping, or reversing the progression of a CD30-expressing disease in asubject.

The terms “specific binding” and “specifically binds” mean that theanti-CD30 antibody will react, in a highly selective manner, with itscorresponding target, CD30, and not with the multitude of otherantigens. Typically, the anti-CD30 antibody binds with an affinity of atleast about 1×10⁻⁷ M, and preferably 1×10⁻⁸ M to 1×10⁻⁹ M, 1×10⁻¹⁰ M,1×10⁻¹¹ M, or 1×10⁻¹² M.

The term “monoclonal antibody” refers to an antibody that is derivedfrom a single cell clone, including any eukaryotic or prokaryotic cellclone, or a phage clone, and not the method by which it is produced.Thus, the term “monoclonal antibody” as used herein is not limited toantibodies produced through hybridoma technology.

The terms “identical” or “percent identity,” in the context of two ormore nucleic acids or polypeptide sequences, refer to two or moresequences or subsequences that are the same or have a specifiedpercentage of nucleotides or amino acid residues that are the same, whencompared and aligned for maximum correspondence. To determine thepercent identity, the sequences are aligned for optimal comparisonpurposes (e.g., gaps can be introduced in the sequence of a first aminoacid or nucleic acid sequence for optimal alignment with a second aminoor nucleic acid sequence). The amino acid residues or nucleotides atcorresponding amino acid positions or nucleotide positions are thencompared. When a position in the first sequence is occupied by the sameamino acid residue or nucleotide as the corresponding position in thesecond sequence, then the molecules are identical at that position. Thepercent identity between the two sequences is a function of the numberof identical positions shared by the sequences (i.e., % identity=#ofidentical positions/total #of positions (e.g., overlappingpositions)×100). In certain embodiments, the two sequences are the samelength.

The term “substantially identical,” in the context of two nucleic acidsor polypeptides, refers to two or more sequences or subsequences thathave at least 70% or at least 75% identity; more typically at least 80%or at least 85% identity; and even more typically at least 90%, at least95%, or at least 98% identity (for example, as determined using one ofthe methods set forth below).

The determination of percent identity between two sequences can beaccomplished using a mathematical algorithm. A preferred, non-limitingexample of a mathematical algorithm utilized for the comparison of twosequences is the algorithm of Karlin and Altschul, 1990, Proc. Natl.Acad. Sci. USA 87:2264-2268, modified as in Karlin and Altschul, 1993,Proc. Natl. Acad. Sci. USA 90:5873-5877. Such an algorithm isincorporated into the NBLAST and XBLAST programs of Altschul, et al.,1990, J. Mol. Biol. 215:403-410. BLAST nucleotide searches can beperformed with the NBLAST program, score=100, wordlength=12 to obtainnucleotide sequences homologous to a nucleic acid encoding a protein ofinterest. BLAST protein searches can be performed with the XBLASTprogram, score=50, wordlength=3 to obtain amino acid sequenceshomologous to protein of interest. To obtain gapped alignments forcomparison purposes, Gapped BLAST can be utilized as described inAltschul et al., 1997, Nucleic Acids Res. 25:3389-3402. Alternatively,PSI-Blast can be used to perform an iterated search which detectsdistant relationships between molecules (Id.). When utilizing BLAST,Gapped BLAST, and PSI-Blast programs, the default parameters of therespective programs (e.g., XBLAST and NBLAST) can be used. Anotherpreferred, non-limiting example of a mathematical algorithm utilized forthe comparison of sequences is the algorithm of Myers and Miller, CABIOS(1989). Such an algorithm is incorporated into the ALIGN program(version 2.0) which is part of the GCG sequence alignment softwarepackage. When utilizing the ALIGN program for comparing amino acidsequences, a PAM120 weight residue table, a gap length penalty of 12,and a gap penalty of 4 can be used. Additional algorithms for sequenceanalysis are known in the art and include ADVANCE and ADAM as describedin Torellis and Robotti, 1994, Comput. Appl. Biosci. 10:3-5; and FASTAdescribed in Pearson and Lipman, 1988, Proc. Natl. Acad. Sci. 85:2444-8.Within FASTA, ktup is a control option that sets the sensitivity andspeed of the search. If ktup=2, similar regions in the two sequencesbeing compared are found by looking at pairs of aligned residues; ifktup=1, single aligned amino acids are examined. ktup can be set to 2 or1 for protein sequences, or from 1 to 6 for DNA sequences. The defaultif ktup is not specified is 2 for proteins and 6 for DNA.

Alternatively, protein sequence alignment may be carried out using theCLUSTAL W algorithm, as described by Higgins et al., 1996, MethodsEnzymol. 266:383-402.

The term “pharmaceutically acceptable” as used herein refers to thosecompounds, materials, compositions, and/or dosage forms that are, withinthe scope of sound medical judgment, suitable for contact with thetissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problems or complicationscommensurate with a reasonable benefit/risk ratio. The term“pharmaceutically compatible ingredient” refers to a pharmaceuticallyacceptable diluent, adjuvant, excipient, or vehicle with which anantibody-drug conjugate is administered.

The abbreviation “MMAE” refers to monomethyl auristatin E.

The abbreviations “vc” and “val-cit” refer to the dipeptidevaline-citrulline.

The abbreviation “PAB” refers to the self-immolative spacer:

The abbreviation “MC” refers to the stretcher maleimidocaproyl:

cAC10-MC-vc-PAB-MMAE refers to a chimeric AC10 antibody conjugated tothe drug MMAE through a MC-vc-PAB linker.

An anti-CD30 vc-PAB-MMAE antibody-drug conjugate refers to an anti-CD30antibody conjugated to the drug MMAE via a linker comprising thedipeptide valine citrulline and the self-immolative spacer PAB as shownin Formula (I).

The phrase “single weekly dose” as it refers to administration of anantibody-drug conjugate refers to administration of the drug once in aweekly period. In contrast, a “split-delivery dose” during a weeklyperiod refers to administration of the drug more than once during theweekly time period.

As used herein, “pharmaceutically acceptable salts” refer to derivativesof the disclosed compounds wherein the parent compound is modified bymaking acid or base salts thereof. Examples of pharmaceuticallyacceptable salts include, but are not limited to, mineral or organicacid salts of basic residues such as amines; alkali or organic salts ofacidic residues such as carboxylic acids; and the like. Thepharmaceutically acceptable salts include the conventional non-toxicsalts or the quaternary ammonium salts of the parent compound formed,for example, from non-toxic inorganic or organic acids. For example,such conventional non-toxic salts include those derived from inorganicacids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric,nitric and the like; and the salts prepared from organic acids such asacetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric,citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic,benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric,toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic,and the like. These physiologically acceptable salts are prepared bymethods known in the art, e.g., by dissolving the free amine bases withan excess of the acid in aqueous alcohol, or neutralizing a freecarboxylic acid with an alkali metal base such as a hydroxide, or withan amine.

Unless otherwise noted, the term “alkyl” refers to a saturated straightor branched hydrocarbon having from about 1 to about 20 carbon atoms(and all combinations and subcombinations of ranges and specific numbersof carbon atoms therein), with from about 1 to about 8 carbon atomsbeing preferred. Examples of alkyl groups are methyl, ethyl, n-propyl,iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, n-pentyl,2-pentyl, 3-pentyl, 2-methyl-2-butyl, n-hexyl, n-heptyl, n-octyl,n-nonyl, n-decyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl,1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl,4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl,2,3-dimethyl-2-butyl, and 3,3-dimethyl-2-butyl.

Alkyl groups, whether alone or as part of another group, can beoptionally substituted with one or more groups, preferably 1 to 3 groups(and any additional substituents selected from halogen), including, butnot limited to, -halogen, —O—(C₁-C₈ alkyl), —O—(C₂-C₈ alkenyl),—O—(C₂-C₈ alkynyl), -aryl, —C(O)R′, —OC(O)R′, —C(O)OR′, —C(O)NH₂,—C(O)NHR′, —C(O)N(R′)₂, —NHC(O)R′, —SR′, —SO₃R′, —S(O)₂R′, —S(O)R′, —OH,═O, —N₃, —NH₂, —NH(R′), —N(R′)₂ and —CN, where each R′ is independentlyselected from —H, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, or-aryl, and wherein said —O—(C₁-C₈ alkyl), —O—(C₂-C₈ alkenyl), —O—(C₂-C₈alkynyl), -aryl, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, and —C₂-C₈ alkynyl groupscan be optionally further substituted with one or more groups including,but not limited to, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl,-halogen, —O—(C₁-C₈ alkyl), —O—(C₂-C₈ alkenyl), —O—(C₂-C₈ alkynyl),-aryl, —C(O)R″, —OC(O)R″, —C(O)OR″, —C(O)NH₂, —C(O)NHR″, —C(O)N(R″)₂,—NHC(O)R″, —SR″, —SO₃R″, —S(O)₂R″, —S(O)R″, —OH, —N₃, —NH₂, —NH(R″),—N(R″)₂ and —CN, where each R″ is independently selected from —H, —C₁-C₈alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, or -aryl.

Unless otherwise noted, the terms “alkenyl” and “alkynyl” refer tostraight and branched carbon chains having from about 2 to about 20carbon atoms (and all combinations and subcombinations of ranges andspecific numbers of carbon atoms therein), with from about 2 to about 8carbon atoms being preferred. An alkenyl chain has at least one doublebond in the chain and an alkynyl chain has at least one triple bond inthe chain. Examples of alkenyl groups include, but are not limited to,ethylene or vinyl, allyl, -1-butenyl, -2-butenyl, -isobutylenyl,-1-pentenyl, -2-pentenyl, -3-methyl-1-butenyl, -2-methyl-2-butenyl, and-2,3-dimethyl-2-butenyl. Examples of alkynyl groups include, but are notlimited to, acetylenic, propargyl, acetylenyl, propynyl, -1-butynyl,-2-butynyl, -1-pentynyl, -2-pentynyl, and -3-methyl-1 butynyl.

Alkenyl and alkynyl groups, whether alone or as part of another group,can be optionally substituted with one or more groups, preferably 1 to 3groups (and any additional substituents selected from halogen),including but not limited to, -halogen, —O—(C₁-C₈ alkyl), —O—(C₂-C₈alkenyl), —O—(C₂-C₈ alkynyl), -aryl, —C(O)R′, —OC(O)R′, —C(O)OR′,—C(O)NH₂, —C(O)NHR′, —C(O)N(R′)₂, —NHC(O)R′, —SR′, —SO₃R′, —S(O)₂R′,—S(O)R′, —OH, ═O, —N₃, —NH₂, —NH(R′), —N(R′)₂ and —CN, where each R′ isindependently selected from —H, —C₁-C₈ alkyl, —C₂-C₈ alkyenl, —C₂-C₈alkynyl, or -aryl and wherein said —O—(C₁-C₈ alkyl), —O—(C₂-C₈ alkenyl),—O—(C₂-C₈ alkynyl), -aryl, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, and —C₂-C₈alkynyl groups can be optionally further substituted with one or moresubstituents including, but not limited to, —C₁-C₈ alkyl, —C₂-C₈alkenyl, —C₂-C₈ alkynyl, -halogen, —O—(C₁-C₈ alkyl), —O—(C₂-C₈ alkenyl),—O—(C₂C₈ alkynyl), -aryl, —C(O)R″, —OC(O)R″, —C(O)OR″, —C(O)NH₂,—C(O)NHR″, —C(O)N(R″)₂, —NHC(O)R″, —SR″, —SO₃R″, —S(O)₂R″, —S(O)R″, —OH,—N₃, —NH₂, —NH(R″), —N(R″)₂ and —CN, where each R″ is independentlyselected from —H, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, or-aryl.

Unless otherwise noted, the term “alkylene” refers to a saturatedbranched or straight chain hydrocarbon radical having from about 1 toabout 20 carbon atoms (and all combinations and subcombinations ofranges and specific numbers of carbon atoms therein), with from about 1to about 8 carbon atoms being preferred and having two monovalentradical centers derived by the removal of two hydrogen atoms from thesame or two different carbon atoms of a parent alkane. Typical alkylenesinclude, but are not limited to, methylene, ethylene, propylene,butylene, pentylene, hexylene, heptylene, ocytylene, nonylene, decalene,1,4-cyclohexylene, and the like. Alkylene groups, whether alone or aspart of another group, can be optionally substituted with one or moregroups, preferably 1 to 3 groups (and any additional substituentsselected from halogen), including, but not limited to, -halogen,—O—(C₁-C₈ alkyl), —O—(C₂-C₈ alkenyl), —O—(C₂-C₈ alkynyl), -aryl,—C(O)R′, —OC(O)R′, —C(O)OR′, —C(O)NH₂, —C(O)NHR′, —C(O)N(R′)₂,—NHC(O)R′, —SR′, —SO₃R′, —S(O)₂R′, —S(O)R′, ═O, —N₃, —NH₂, —NH(R′),—N(R′)₂ and —CN, where each R′ is independently selected from —H, —C₁-C₈alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, or -aryl and wherein said—O—(C₁-C₈ alkyl), —O—(C₂-C₈ alkenyl), —O—(C₂-C₈ alkynyl), -aryl, —C₁-C₈alkyl, —C₂-C₈ alkenyl, and —C₂-C₈ alkynyl groups can be furtheroptionally substituted with one or more substituents including, but notlimited to, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, -halogen,—O—(C₁-C₈ alkyl), —O—(C₂-C₈ alkenyl), —O—(C₂-C₈ alkynyl), -aryl,—C(O)R″, —OC(O)R″, —C(O)OR″, —C(O)NH₂—C(O)NHR″, —C(O)N(R″)₂, —NHC(O)R″,—SR″, —SO₃R″, —S(O)₂R″, —S(O)R″, —OH, —N₃, —NH₂, —NH(R″), —N(R″)₂ and—CN, where each R″ is independently selected from —H, —C₁-C₈ alkyl,—C₂-C₈ alkenyl, —C₂-C₈ alkynyl, or -aryl.

Unless otherwise noted, the term “alkenylene” refers to an optionallysubstituted alkylene group containing at least one carbon-carbon doublebond. Exemplary alkenylene groups include, for example, ethenylene(—CH═CH—) and propenylene (—CH═CHCH₂—).

Unless otherwise noted, the term “alkynylene” refers to an optionallysubstituted alkylene group containing at least one carbon-carbon triplebond. Exemplary alkynylene groups include, for example, acetylene(—C≡C—), propargyl (—CH₂C≡C—), and 4-pentynyl (—CH₂CH₂CH₂C≡CH—).

Unless otherwise noted, the term “aryl” refers to a monovalent aromatichydrocarbon radical of 6-20 carbon atoms (and all combinations andsubcombinations of ranges and specific numbers of carbon atoms therein)derived by the removal of one hydrogen atom from a single carbon atom ofa parent aromatic ring system. Some aryl groups are represented in theexemplary structures as “Ar”. Typical aryl groups include, but are notlimited to, radicals derived from benzene, substituted benzene, phenyl,naphthalene, anthracene, biphenyl, and the like.

An aryl group, whether alone or as part of another group, can beoptionally substituted with one or more, preferably 1 to 5, or even 1 to2 groups including, but not limited to, -halogen, —C₁-C₈ alkyl, —C₂-C₈alkenyl, —C₂-C₈ alkynyl, —O—(C₁-C₈ alkyl), —O—(C₂-C₈ alkenyl), —O—(C₂-C₈alkynyl), -aryl, —C(O)R′, —OC(O)R′, —C(O)OR′, —C(O)NH₂, —C(O)NHR′,—C(O)N(R′)₂, —NHC(O)R′, —SR′, —SO₃R′, —S(O)₂R′, —S(O)R′, —OH, —NO₂, —N₃,—NH₂, —NH(R′), —N(R′)₂ and —CN, where each R′ is independently selectedfrom —H, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, or -aryl andwherein said —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, O—(C₁-C₈alkyl), —O—(C₂-C₈ alkenyl), —O—(C₂-C₈ alkynyl), and -aryl groups can befurther optionally substituted with one or more substituents including,but not limited to, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl,-halogen, —O—(C₁-C₈ alkyl), —O—(C₂-C₈ alkenyl), —O—(C₂-C₈ alkynyl),-aryl, —C(O)R″, —OC(O)R″, —C(O)OR″, —C(O)NH₂, —C(O)NHR″, —C(O)N(R″)₂,—NHC(O)R″, —SR″, —SO₃R″, —S(O)₂R″, —S(O)R″, —OH, —N₃, —NH₂, —NH(R″),—N(R″)₂ and —CN, where each R″ is independently selected from —H, —C₁-C₈alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, or -aryl.

Unless otherwise noted, the term “arylene” refers to an optionallysubstituted aryl group which is divalent (i.e., derived by the removalof two hydrogen atoms from the same or two different carbon atoms of aparent aromatic ring system) and can be in the ortho, meta, or paraconfigurations as shown in the following structures with phenyl as theexemplary aryl group:

Typical “—(C₁-C₈ alkylene)aryl,” “—(C₂-C₈ alkenylene)aryl”, “and —(C₂-C₈alkynylene)aryl” groups include, but are not limited to, benzyl,2-phenylethan-1-yl, 2-phenylethen-1-yl, naphthylmethyl,2-naphthylethan-1-yl, 2-naphthylethen-1-yl, naphthobenzyl,2-naphthophenylethan-1-yl and the like.

Unless otherwise noted, the term “heterocycle,” refers to a monocyclic,bicyclic, or polycyclic ring system having from 3 to 14 ring atoms (alsoreferred to as ring members) wherein at least one ring atom in at leastone ring is a heteroatom selected from N, O, P, or S (and allcombinations and subcombinations of ranges and specific numbers ofcarbon atoms and heteroatoms therein). The heterocycle can have from 1to 4 ring heteroatoms independently selected from N, O, P, or S. One ormore N, C, or S atoms in a heterocycle can be oxidized. A monocylicheterocycle preferably has 3 to 7 ring members (e.g., 2 to 6 carbonatoms and 1 to 3 heteroatoms independently selected from N, O, P, or S),and a bicyclic heterocycle preferably has 5 to 10 ring members (e.g., 4to 9 carbon atoms and 1 to 3 heteroatoms independently selected from N,O, P, or S). The ring that includes the heteroatom can be aromatic ornon-aromatic. Unless otherwise noted, the heterocycle is attached to itspendant group at any heteroatom or carbon atom that results in a stablestructure.

Heterocycles are described in Paquette, “Principles of ModernHeterocyclic Chemistry” (W. A. Benjamin, New York, 1968), particularlyChapters 1, 3, 4, 6, 7, and 9; “The Chemistry of Heterocyclic Compounds,A series of Monographs” (John Wiley & Sons, New York, 1950 to present),in particular Volumes 13, 14, 16, 19, and 28; and J. Am. Chem. Soc.82:5566 (1960).

Unless otherwise noted, the term “heterocyclo” refers to an optionallysubstituted heterocycle group as defined above that is divalent (i.e.,derived by the removal of two hydrogen atoms from the same or twodifferent carbon atoms of a parent heterocyclic ring system).

Examples of “heterocycle” groups include by way of example and notlimitation pyridyl, dihydropyridyl, tetrahydropyridyl (piperidyl),thiazolyl, pyrimidinyl, furanyl, thienyl, pyrrolyl, pyrazolyl,imidazolyl, tetrazolyl, benzofuranyl, thianaphthalenyl, indolyl,indolenyl, quinolinyl, isoquinolinyl, benzimidazolyl, piperidinyl,4-piperidonyl, pyrrolidinyl, 2-pyrrolidonyl, pyrrolinyl,tetrahydrofuranyl, bis-tetrahydrofuranyl, tetrahydropyranyl,bis-tetrahydropyranyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl,decahydroquinolinyl, octahydroisoquinolinyl, azocinyl, triazinyl,6H-1,2,5-thiadiazinyl, 2H,6H-1,5,2-dithiazinyl, thienyl, thianthrenyl,pyranyl, isobenzofuranyl, chromenyl, xanthenyl, phenoxathinyl,2H-pyrrolyl, isothiazolyl, isoxazolyl, pyrazinyl, pyridazinyl,indolizinyl, isoindolyl, 3H indolyl, 1H-indazolyl, purinyl, 4Hquinolizinyl, phthalazinyl, naphthyridinyl, quinoxalinyl, quinazolinyl,cinnolinyl, pteridinyl, 4H-carbazolyl, carbazolyl, β-carbolinyl,phenanthridinyl, acridinyl, pyrimidinyl, phenanthrolinyl, phenazinyl,phenothiazinyl, furazanyl, phenoxazinyl, isochromanyl, chromanyl,imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, piperazinyl,indolinyl, isoindolinyl, quinuclidinyl, morpholinyl, oxazolidinyl,benzotriazolyl, benzisoxazolyl, oxindolyl, benzoxazolinyl, andisatinoyl. Preferred “heterocycle” groups include, but are not limitedto, benzofuranyl, benzothiophenyl, indolyl, benzopyrazolyl, coumarinyl,isoquinolinyl, pyrrolyl, thiophenyl, furanyl, thiazolyl, imidazolyl,pyrazolyl, triazolyl, quinolinyl, pyrimidinyl, pyridinyl, pyridonyl,pyrazinyl, pyridazinyl, isothiazolyl, isoxazolyl and tetrazolyl.

A heterocycle group, whether alone or as part of another group, can beoptionally substituted with one or more groups, preferably 1 to 2groups, including but not limited to, —C₁-C₈ alkyl, —C₂-C₈ alkenyl,—C₂-C₈ alkynyl, -halogen, —O—(C₁-C₈ alkyl), —O—(C₂-C₈ alkenyl),—O—(C₂-C₈ alkynyl), -aryl, —C(O)R′, —OC(O)R′, —C(O)OR′, —C(O)NH₂,—C(O)NHR′, —C(O)N(R′)₂, —NHC(O)R′, —SR′, —SO₃R′, —S(O)₂R′, —S(O)R′, —OH,—N₃, —NH₂, —NH(R′), —N(R′)₂ and —CN, where each R′ is independentlyselected from —H, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, alkynyl, or -aryl andwherein said —O—(C₁-C₈ alkyl), —O—(C₂-C₈ alkenyl), —O—(C₂-C₈ alkynyl),—C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, and -aryl groups can befurther optionally substituted with one or more substituents including,but not limited to, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl,-halogen, —O—(C₁-C₈ alkyl), —O—(C₂-C₈ alkenyl), —O—(C₂-C₈ alkynyl),-aryl, —C(O)R″, —OC(O)R″, —C(O)OR″, —C(O)NH₂, —C(O)NHR″, —C(O)N(R″)₂,—NHC(O)R″, —SR″, —SO₃R″, —S(O)₂R″, —S(O)R″, —OH, —N₃, —NH₂, —NH(R″),—N(R″)₂ and —CN, where each R″ is independently selected from —H, —C₁-C₈alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, or aryl.

Unless otherwise noted, the term “carbocycle,” refers to a saturated orunsaturated non-aromatic monocyclic, bicyclic, or polycyclic ring systemhaving from 3 to 14 ring atoms (and all combinations and subcombinationsof ranges and specific numbers of carbon atoms therein) wherein all ofthe ring atoms are carbon atoms. Monocyclic carbocycles preferably have3 to 6 ring atoms, still more preferably 5 or 6 ring atoms. Bicycliccarbocycles preferably have 7 to 12 ring atoms, e.g., arranged as abicyclo [4,5], [5,5], [5,6] or [6,6] system, or 9 or 10 ring atomsarranged as a bicyclo [5,6] or [6,6] system. The term “carbocycle”includes, for example, a monocyclic carbocycle ring fused to an arylring (e.g., a monocyclic carbocycle ring fused to a benzene ring).Carbocyles preferably have 3 to 8 carbon ring atoms.

Carbocycle groups, whether alone or as part of another group, can beoptionally substituted with, for example, one or more groups, preferably1 or 2 groups (and any additional substituents selected from halogen),including, but not limited to, -halogen, —C₁-C₈ alkyl, —C₂-C₈ alkenyl,—C₂-C₈ alkynyl, —O—(C₁-C₈ alkyl), —O—(C₂-C₃ alkenyl), —O—(C₂-C₈alkynyl), -aryl, —C(O)R′, —OC(O)R′, —C(O)OR′, —C(O)NH₂—C(O)NHR′,—C(O)N(R′)₂, —NHC(O)R′, —SR′, —SO₃R′, —S(O)₂R′, —S(O)R′, —OH, ═O, —N₃,—NH₂, —NH(R′), —N(R′)₂ and —CN, where each R′ is independently selectedfrom —H, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, or -aryl andwherein said —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, —O—(C₁-C₈alkyl), —O—(C₂-C₈ alkenyl), —O—(C₂-C₈ alkynyl), and -aryl groups can befurther optionally substituted with one or more substituents including,but not limited to, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl,-halogen, —O—(C₁-C₈ alkyl), —O—(C₂-C₈ alkenyl), —O—(C₂-C₈ alkynyl),-aryl, —C(O)R″, —OC(O)R″, —C(O)OR″, —C(O)NH₂, —C(O)NHR″, —C(O)N(R″)₂,—NHC(O)R″, —SR″, —SO₃R″, —S(O)₂R″, —S(O)R″, —OH, —N₃, —NH₂, —NH(R″),—N(R″)₂ and —CN, where each R″ is independently selected from —H, —C₁-C₈alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, or -aryl.

Examples of monocyclic carbocylic substituents include -cyclopropyl,-cyclobutyl, -cyclopentyl, -1-cyclopent-1-enyl, -1-cyclopent-2-enyl,-1-cyclopent-3-enyl, cyclohexyl, -1-cyclohex-1-enyl, -1-cyclohex-2-enyl,-1-cyclohex-3-enyl, -cycloheptyl, -cyclooctyl, -1,3-cyclohexadienyl,-1,4-cyclohexadienyl, -1,3-cycloheptadienyl, -1,3,5-cycloheptatrienyl,and -cyclooctadienyl.

A “carbocyclo,” whether used alone or as part of another group, refersto an optionally substituted carbocycle group as defined above that isdivalent (i.e., derived by the removal of two hydrogen atoms from thesame or two different carbon atoms of a parent carbocyclic ring system).

Unless otherwise indicated by context, a hyphen (-) designates the pointof attachment to the pendant molecule. Accordingly, the term “—(C₁-C₈alkylene)aryl” or “—C₁-C₈ alkylene(aryl)” refers to a C₁-C₈ alkyleneradical as defined herein wherein the alkylene radical is attached tothe pendant molecule at any of the carbon atoms of the alkylene radicaland one of the hydrogen atoms bonded to a carbon atom of the alkyleneradical is replaced with an aryl radical as defined herein.

When a particular group is “substituted”, that group may have one ormore substituents, preferably from one to five substituents, morepreferably from one to three substituents, most preferably from one totwo substituents, independently selected from the list of substituents.Groups that are substituted are so indicated.

It is intended that the definition of any substituent or variable at aparticular location in a molecule be independent of its definitionselsewhere in that molecule. It is understood that substituents andsubstitution patterns on the compounds of this invention can be selectedby one of ordinary skill in the art to provide compounds that arechemically stable and that can be readily synthesized by techniquesknown in the art as well as those methods set forth herein.

D. Dosing Regimen

The present invention provides, inter alia, a dosing regimen for thetreatment of CD30-expressing hematologic cancers. The dosing regimencomprises a weekly dose of an antibody-drug conjugate as describedherein of from about 0.8 mg/kg body weight to about 1.8 mg/kg bodyweight, 0.8 mg/kg body weight to about 1.6 mg/kg body weight, 0.8 mg/kgbody weight to about 1.4 mg/kg body weight, 0.8 mg/kg body weight toabout 1.2 mg/kg body weight, or more preferably from about 0.8 mg/kgbody weight to about 1.0 mg/kg body weight for at least a three weekperiod (e.g., 21 day period). The weekly dose can either be administeredas a single weekly dose (once a week) or by split delivery (e.g., two ormore times per week).

In some embodiments, the weekly dose of the antibody drug conjugate willbe about 0.8 mg/kg body weight. In some embodiments, the weekly dose ofthe antibody drug conjugate will be about 0.9 mg/kg body weight. In someembodiments, the weekly dose of the antibody drug conjugate will beabout 1.0 mg/kg body weight. In some embodiments, the weekly dose of theantibody drug conjugate will be about 1.1 mg/kg body weight. In someembodiments, the weekly dose of the antibody drug conjugate will beabout 1.2 mg/kg body weight. In some embodiments, the weekly dose of theantibody drug conjugate will be about 1.3 mg/kg body weight. In someembodiments, the weekly dose of the antibody drug conjugate will beabout 1.4 mg/kg body weight. In some embodiments, the weekly dose of theantibody drug conjugate will be about 1.5 mg/kg body weight. In someembodiments, the weekly dose of the antibody drug conjugate will beabout 1.6 mg/kg body weight. In some embodiments, the weekly dose of theantibody drug conjugate will be about 1.7 mg/kg body weight. In someembodiments, the weekly dose of the antibody drug conjugate will beabout 1.8 mg/kg body weight. In some embodiments, the weekly dose of theantibody drug conjugate will be 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5,1.6, 1.7 or 1.8 mg/kg of the subject's body weight.

In some embodiments, the weekly dose is administered, as a splitdelivery or as a single weekly dose, for at least one three week (e.g.,21 day) treatment cycle. In some embodiments, the dose will beadministered as a single weekly dose on days 1, 8, and 15 of a 21 daytreatment cycle. Preferably, the weekly dose, as a split delivery or asa single weekly dose, is administered for two or more 21 day treatmentcycles, even more preferably for three or more, four or more, five, oreven six or more treatment cycles. In some embodiments, the weekly doseis administered for no more than 3, no more than 4, no more than 5, orno more than 6 treatment cycles. Preferably, there will a period of restbetween treatment cycles. For example, in some preferred embodiments,the dosing regimen will be a total weekly dose of the antibody-drugconjugate of from about 0.8 mg/kg body weight to about 1.8 mg/kg bodyweight, 0.8 mg/kg body weight to about 1.6 mg/kg body weight, 0.8 mg/kgbody weight to about 1.4 mg/kg body weight, 0.8 mg/kg body weight toabout 1.2 mg/kg body weight or 0.8 mg/kg body weight to about 1.0 mg/kgbody weight for at least two treatment cycles with a one week period ofrest between each of the treatment cycles (e.g., six single weekly dosesduring an eight week time period). In some embodiments, the treatmentcycle will be greater than 21 days. The weekly dose can be administeredas a single weekly dose (once a week) or by split delivery (e.g., two ormore times per week).

Instead of referring to the treatment cycle as a three week treatmentcycle with a one week period of rest between treatment cycles, thetreatment cycle can be referred to as a 4 week (28 day) treatment cyclewhere the antibody-drug conjugate is delivered 3 out of 4 weeks in the 4week treatment cycle. Accordingly, in some embodiments, the dose isadministered weekly, as a split delivery or as a single weekly dose, 3out of 4 weeks in a 4 week treatment cycle. In some embodiments, thedose will be administered as a single weekly dose on days 1, 8, and 15of a 28 day treatment cycle. Preferably, the weekly dose, as a splitdelivery or as a single weekly dose, is administered for two or morefour week treatment cycles, even more preferably for three or more, fouror more, five or more, or even six or more four week treatment cycles(e.g., 2, 3, 4, 5, or 6 consecutive treatment cycles). In someembodiments, the weekly dose is administered for no more than 3, no morethan 4, no more than 5, or no more than 6 treatment cycles. For example,in some preferred embodiments, the dosing regimen will be a weekly dose,as a split delivery or as a single weekly dose, for a total weekly doseof from about 0.8 mg/kg body weight to about 1.8 mg/kg body weight, 0.8mg/kg body weight to about 1.6 mg/kg body weight, 0.8 mg/kg body weightto about 1.4 mg/kg body weight, 0.8 mg/kg body weight to about 1.2 mg/kgbody weight or 0.8 mg/kg body weight to about 1.0 mg/kg body weight ofthe antibody-drug conjugate, 3 out of 4 weeks, for at least two fourweek treatment cycles (e.g., six single weekly doses an eight week timeperiod). In some preferred embodiments, the dosing regimen will be aweekly dose, as a split delivery or as a single weekly dose, for a totalweekly dose of from about 0.8 mg/kg body weight to about 1.8 mg/kg bodyweight, 0.8 mg/kg body weight to about 1.6 mg/kg body weight, 0.8 mg/kgbody weight to about 1.4 mg/kg body weight, 0.8 mg/kg body weight toabout 1.2 mg/kg body weight or 0.8 mg/kg body weight to about 1.0 mg/kgbody weight of the antibody-drug conjugate, 3 out of 4 weeks, for one,two, three, four, or five four week treatment cycles (e.g., six singleweekly doses in an eight week time period, nine single weekly doses in atwelve week time period, twelve single weekly doses in a sixteen weektime period).

Following or during one or more treatment cycles (e.g., during days21-28 of the second treatment cycle), the subject can be evaluated(e.g., through clinical or diagnostic testing) to determine whether thesubject should remain on the treatment schedule. For example, followingor during one or more 28 day treatment cycles (e.g., 1, 2, 3, 4, 5, or 628 day treatment cycles), the subject can be evaluated (e.g., a clinicaland/or diagnostic evaluation). Depending on the evaluation, the subjectwill discontinue treatment, continue on treatment with additionaltreatment cycles, or commence maintenance therapy. If the subjectcontinues treatment, the subject can be further evaluated following oneor more additional treatment cycles. Depending on each successiveevaluation, the subject will discontinue treatment, continue ontreatment with additional treatment cycles, or commence maintenancetherapy.

The present invention encompasses embodiments wherein the subjectremains on the weekly treatment cycle (e.g., the four week treatmentcycle) following an evaluation indicating that the subject has nodetectable cancer, for example, following a diagnostic test that isnegative for the CD30 expressing cancer (i.e., the diagnostic test isunable to detect any cancer in the subject). For example, in someembodiments, the subject will remain on the weekly treatment cycle forat least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more treatment cyclesfollowing such an evaluation. In some embodiments, the subject willremain on the weekly treatment cycle for at least two but no more than3, no more than 4, no more than 5, or no more than 6 treatment cycles.One example of a diagnostic test used for determining the presence andseverity of cancers is positron emission tomography (PET).

In some embodiments, the subject will commence maintenance therapyfollowing one or more, preferably two or more, (e.g, following 1, 2, 3,4, 5, or 6) treatment cycles (e.g., the four week treatment cycle). Insome embodiments, the subject will commence maintenance therapyfollowing an evaluation indicating that the subject has little or nodetectable cancer, e.g., following an evaluation indicating that thesubject has had a complete response. As used herein, maintenance therapyrefers to therapy with the antibody-drug conjugate but at a reducedadministration schedule at either the same or different dosages. Duringmaintenance therapy, the antibody-drug conjugate is preferablyadministered once every two weeks, once every three weeks, once everyfour weeks, once every five weeks, once every 6 weeks, once every 7weeks, or once every 8 weeks. Following these maintenance therapycycles, the subject can be further evaluated (e.g., through clinical ordiagnostic testing) to determine whether the subject should remain onthe maintenance therapy, continue with regular treatment or discontinuetreatment. In some embodiments, maintenance therapy will be once everythree weeks to six weeks. The dosage of the antibody drug conjugateadministered during maintenance therapy can range, for example, fromabout 0.3 mg/kg body weight to about 2.0 mg/kg body weight, preferablyfrom about 0.6 mg/kg body weight to about 1.8 mg/kg body weight,preferably from about 1.2 mg/kg body weight to about 2.0 mg/kg bodyweight, more preferably from about 1 mg/kg body weight to about 1.8mg/kg body weight per dose, with 1.8 mg/kg being an exemplary dose.

In some embodiments, following conclusion of the weekly treatment at adosage of the antibody drug conjugate of from about 0.8 mg/kg bodyweight to about 1.8 mg/kg body weight, more preferably a dosage of fromabout 0.8 mg/kg body weight to about 1.2 mg/kg body weight andevaluation, the subject will begin a maintenance therapy which comprisesadministration of the antibody-drug conjugate once every three to sixweeks at a dosage of from about 0.3 mg/kg body weight to about 2 mg/kgbody weight, preferably from about 0.6 mg/kg body weight to about 1.8mg/kg body weight, preferably from about 1.2 mg/kg body weight to about2.0 mg/kg body weight, more preferably from about 1 mg/kg body weight toabout 1.8 mg/kg body weight with about 1.8 mg/kg being preferred. Insome embodiments, following conclusion of the weekly treatment (e.g.,for one, two, three, four or five treatment cycles), the subject willbegin a once every three week administration schedule (e.g., treatmenton day 1 of a three week maintenance therapy cycle) of the antibody drugconjugate at a dosage of from about 0.4 mg/kg body weight to about 2mg/kg body weight, from about 1.2 mg/kg body weight to about 2.0 mg/kgbody weight, or from about 1 mg/kg body weight to about 1.8 mg/kg bodyweight with about 1.8 mg/kg being preferred.

Accordingly, the present invention encompasses embodiments wherein asubject will be administered a weekly dose, as a split delivery or as asingle weekly dose, of the antibody drug conjugate for a total weeklydose of from about 0.8 mg/kg of the subject's body weight to about 1.8mg/kg of the subject's body weight, about 0.8 mg/kg body weight to about1.6 mg/kg body weight, about 0.8 mg/kg body weight to about 1.4 mg/kgbody weight, about 0.8 mg/kg body weight to about 1.2 mg/kg body weightor about 0.8 mg/kg body weight to about 1.0 mg/kg body weight, 3 out of4 weeks, for one, two, three, four, five, or six 28 day treatment cyclesfollowed by administration of an every three to six week dose,preferably every three week dose, of antibody drug conjugate at a doseof from about 0.4 mg/kg body weight to about 2 mg/kg body weight, fromabout 1.2 mg/kg body weight to about 2.0 mg/kg body weight, or fromabout 1.0 mg/kg body weight to about 1.8 mg/kg body weight per dose for2 or more maintenance therapy cycles. In some embodiments, the weeklyadministration cycle will be for 2, 3, 4, 5, 6, 7, 8, 9 or 10 or moretreatment cycles and the every three week administration schedule willbe for 2, 3, 4, 5, 6, 7, 8, 9 or 10 or more maintenance therapy cycles.In some embodiments, the weekly administration cycle will be for no morethan 2, 3, 4, 5, or 6 treatment cycles.

The present invention encompasses embodiments wherein a subject will beadministered a weekly dose, as a split delivery or as a single weeklydose, of the antibody drug conjugate at a total weekly dose of dose offrom about 0.8 mg/kg of the subject's body weight to about 1.2 mg/kg 3out of 4 weeks (e.g., on days 1, 8, and 15 of a 28 day treatment cycle)for one, two, three, four, five, or six 28 day treatment cycles followedby administration of an every three to six week dose, preferably everythree week dose, of antibody drug conjugate at a dose of about 1.8 mg/kgper body weight for 2 or more maintenance therapy cycles (e.g., a doseof about 1.8 mg/kg per body weight every three weeks for two or morethree week maintenance therapy cycles).

The present invention encompasses embodiments wherein the subject to betreated by the present methods is being treated with a anti-CD30 vc-PABMMAE antibody-drug conjugate of the present invention but at a scheduleother than the weekly dosing regimen (e.g., administration of theantibody drug conjugate at a dose of about 1.8 mg/kg body weight everythree weeks for one or more three week therapy cycles) and is switchedto a weekly dosing regimen as described herein for no more than 1, 2, 3,4, 5, or 6 treatment cycles. Following the weekly dosing regimen, thepatient can optionally commence maintenance therapy as described herein.

The present invention also encompasses embodiments wherein followingtreatment with the antibody-drug conjugate and prior to commencement ofmaintenance therapy, the subject will undergo a stem cell transplant.Accordingly, in some embodiments, following two or more treatment cycles(e.g., following 1, 2, 3, 4, 5 or 6 treatment cycles) and following anevulation indicating that the subject is eligible for a stem celltransplant, the subject will undergo a stem cell transplant. Followingthe transplant, the subject can commence maintenance therapy.Maintenance therapy will commence post transplant and prior to anydetectable relapse of the cancer.

The antibody-drug conjugate is preferably administered as a monotherapy.By the term “monotherapy” it is meant that the antibody drug conjugateis the only anti-cancer agent administered to the subject during thetreatment cycle. Other therapeutic agents, however, can be administeredto the subject. For example, anti-inflammatory agents or other agentsadministered to a subject with cancer to treat symptoms associated withcancer, but not the underlying cancer itself, including, for exampleinflammation, pain, weight loss, and general malaise can be administeredduring the period of monotherapy. A subject being treated by the presentmethods will preferably have completed any prior treatment withanti-cancer agents before administration of the antibody drug conjugate.In some embodiments, the subject will have completed any prior treatmentwith anti-cancer agents at least 1 week (preferably 2, 3, 4, 5, 6, 7, or8 weeks) prior to treatment with the antibody drug conjugate. Thesubject will also, preferably, not be treated with any additionalanti-cancer agents for at least 2 weeks (preferably at least 3, 4, 5, 6,7, or 8 weeks) following completion of the first treatment cycle withthe antibody drug conjugate and preferably for at least 2 weeks(preferably at least 3, 4, 5, 6, 7, or 8 weeks) following completion ofthe last dose of the antibody drug conjugate.

E. Subjects

The methods of the present invention encompass administering ananti-CD30 vc-PAB-MMAE antibody-drug conjugate to a subject for thetreatment of a CD30-expressing hematologic cancer. After administrationof the antibody-drug conjugate to a subject and binding of the anti-CD30antibody to a CD30 expressing cancer cell, the antibody drug conjugateinternalizes into the cell, and the drug is released.

A CD30-expressing hematologic cancer refers to a hematologic cancer thatexpresses the CD30 antigen. The CD30 antigen is expressed in largenumbers on tumor cells of select lymphomas and leukemias. Hodgkinlymphoma, anaplastic large-cell lymphoma, and/or acute or lymphomatousforms of adult T-cell leukemia are examples of CD30-expressinghematologic cancers that can be treated by the present methods.

The subjects to be treated with the methods of the present invention arethose that have been diagnosed with a CD30 expressing hematologic canceror are suspected of having a CD30-expressing hematologic cancer.Diagnosis can be by methods known in the art, including, for example,lymph node biopsy. After Hodgkin lymphoma is diagnosed, for example, asubject can be classified according to stage of disease using one of theknown classification schemes. The Cotswolds staging classificationscheme is one such classification scheme. The methods of the presentinvention can be used to treat a subject classified in any stage ofdisease, including a subject with advanced stage disease.

The methods of the present invention encompass treating a subject who isnewly diagnosed and has not previously been treated for aCD30-expressing hematologic cancer.

The methods of the present invention can be used to treat subjects witha refractory and/or relapsed CD30-expressing hematologic cancer.

A subject with a refractory CD30-expressing hematologic cancer is asubject who has not responded to a previous anti-cancer therapy, i.e.,the subject continues to experience disease progresssion despitetherapy.

A subject with a relapsed CD30-expressing hematologic cancer is asubject who has responded to a prior anti-cancer therapy for the diseaseat one point, but has had a reoccurence or further progression ofdisease following the response.

The methods of the present invention encompass, for example, treating asubject who has previously been treated with a first-line chemotherapyregimen and/or a salvage regimen and/or experimental treatment for theCD30-expressing hematologic cancer. First line chemotherapeutic regimensfor Hodgkin lymphoma include, for example, the ABVD regimen (Bonadonnaand Santoro, Cancer Treat Rev 1982; 9:21-35), the BEACOPP regimen (Diehlet al., N Engl J Med 2003; 348:2386-2395), the escalated BEACOPP regimen(Diehl et al., N Engl J Med 2003; 348:2386-2395), the MOPP regimen(Devita et al., Ann Inter Med 1970:73:881-895), and the Stanford Vregimen (Horning et al., J Clin Oncol 2000; 18:972-980). Salvagechemotherapy regimens and experimental regimens include, for example,the ESHAP regimen (Aparicio et al., Ann Ocol 1999; 10:593-595), themodified Stanford V regimen (Aviles et al., Med Oncol 2001; 18:261-267),the GDP regimen (Baetz et al., Ann Oncol 2003; 14:1762-1767), theMini-Beam regimen (Colwill et al., J Clin Oncol 1995; 13:396-402,Fernandez-Jimenez et al., Haematologica 1999; 84:1007-1011), the MIMEregimen (Enblad et al., Eur J Haematol 1998; 60:166-171), the MINEregimen (Ferme et al., Ann Oncol 1995; 6:543-549), the IEE regimen(Jackson et al., Leuk Lymphoma 2000; 37:561-570), the DHAP regimen(Josting et al., Ann Oncol 2002; 13:1628-1635), the ICE regimen(Moskowitz et al., Semin Oncol 2004; 31(suppl):54-59), the HVP regimen(Oyan et al., Biol Blood Marrow Transplant 2005; 11:688-697), the IVEregimen (Proctor et al., Eur J Haematol 2001; 64(suppl):28-32), the VIPregimen (Ribrag et al., Bome Marrow Transplant 1998; 21:969-974), theASHAP regimen (Rodriguez et al., Blood 1999; 93:3632-3636), theDexa-BEAM regimen (Schmitz et al., Lancet 2002; 359:2065-2071), the CEPregimen (Szanto et al., Oncology 1991; 48:456-458), the CN30P regimen(Walewski et al., Med Oncol. 2000; 17:195-202), the MVC regimen (Wierniket al., Cancer J Sci Am 1998; 4:254-260) and gemcitabine (Savage et al.,Annals of Oncology 2000; 11:595-597).

The methods of the present invention also encompass treating a subjectwho has previously undergone a stem cell transplant. In someembodiments, the methods encompass treating a subject who has undergonea stem cell transplant for treatment of a CD30-expressing hematologiccancer and who, at the time, of treatment has no detectable signs ofcancer. The methods also encompass treating a subject who has previouslyundergone a stem cell transplant but has relapsed.

F. Anti-CD30 Antibodies

Anti-CD30 antibodies suitable for use in accordance with the presentcompositions and methods include any antibody that specifically binds tothe CD30 antigen.

In some embodiments, anti-CD30 antibodies of the present invention notonly immunospecifically bind CD30 but also can exert cytostatic and/orcytotoxic effect on cancerous cells, for example, malignant cells in HL.In some such embodiments, the cytostatic or cytotoxic effect iscomplement-independent and can be achieved in the absence of (i)conjugation to a cytostatic or cytotoxic agent and (ii) effector cells.For example, in some such embodiments, the cytostatic or cytotoxiceffect is not a result of antibody effector function but, for example, aresult of signaling activity.

Anti-CD30 antibodies of the present invention are preferably monoclonaland can include, for example, chimeric (e.g., having a human constantregion and mouse variable region), humanized, or human antibodies. Theimmunoglobulin molecule is of the IgG type and can be any subclass(e.g., IgG1, IgG2, IgG3, IgG4) of immunoglobulin molecule and variantsthereof. The immunoglobulin molecule is preferably an IgG1.

The antibodies of the present invention can be generated by any suitablemethod known in the art. Monoclonal antibodies can be prepared using awide variety of techniques known in the art including the use ofhybridoma, recombinant, and phage display technologies, or a combinationthereof. For example, monoclonal antibodies can be produced usinghybridoma techniques including those known in the art and taught, forexample, in Harlow et al., Antibodies: A Laboratory Manual, (Cold SpringHarbor Laboratory Press, 2nd ed., 1988); Hammerling, et al., in:Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier, N.Y.,1981) (said references incorporated by reference in their entireties).

Any method known in the art for the synthesis of proteins, e.g.,recombinant expression techniques, can be used to generate the anti-CD30antibodies of the present invention.

Once a CD30-binding protein is identified, if desired, its ability(alone or when multimerized or fused to a dimerization ormultimerization domain) to elicit a cytostatic or cytotoxic effect onCD30-expressing cancerous cells can be determined by contacting aculture of an CD30-expressing cancer cell line, such as L428, L450,HLLM2 or KM-H2, with the protein. Culture conditions are most preferablyabout 5,000 cells in a culture area of about 0.33 cm², and thecontacting period being approximately 72 hours. The culture is thenexposed to 0.5 μCi of ³H-thymidine during the final 8 hours of the72-hour period and the incorporation of ³H-thymidine into cells of theculture is measured. The protein has a cytostatic or cytotoxic effect onthe cell line if the cells of the culture have reduced ³H-thymidineincorporation compared to cells of the same cell line cultured under thesame conditions but not contacted with the protein. There are many othercytotoxicity assays known to those of skill in the art. Any one of themcan be used in the present methods.

Exemplary anti-CD30 antibodies include, but are not limited to,humanized or chimeric AC10 or HeFi-1 antibodies. Murine AC10 has beendeposited under ATCC Accession Number PTA-6679.

An exemplary anti-CD30 antibody comprises one or more (1, 2, 3, 4, 5, or6) CDRs of murine HeFi-1 (SEQ ID NO:20, SEQ ID NO:22; SEQ ID NO:24; SEQID NO:28, SEQ ID NO:30 or SEQ ID NO:32) or murine AC10 (SEQ ID NO:4; SEQID NO:6; SEQ ID NO:8; SEQ ID NO:12; SEQ ID NO:14; or SEQ ID NO:16). Insome embodiments, the anti-CD30 antibody comprises the heavy and/orlight chain variable regions of murine HcFi-1 (SEQ ID NO:18 or SEQ IDNO:26) or murine AC10 (SEQ ID NO:2 or SEQ ID NO:10). A table indicatingthe region of AC10 or HeFi-1 to which each SEQ ID NO corresponds to isprovided below.

TABLE 1 NUCLEOTIDE OR MOLECULE AMINO ACID SEQ ID NO AC10 Heavy ChainVariable Region Nucleotide 1 AC10 Heavy Chain Variable Region Amino Acid2 AC10 Heavy Chain-CDR1(H1) Nucleotide 3 AC 10 Heavy Chain-CDR1(H1)Amino Acid 4 AC 10 Heavy Chain-CDR2(H2) Nucleotide 5 AC 10 HeavyChain-CDR2(H2) Amino Acid 6 AC 10 Heavy Chain-CDR3(H3) Nucleotide 7 AC10 Heavy Chain-CDR3(H3) Amino Acid 8 AC 10 Light Chain Variable RegionNucleotide 9 AC 10 Light Chain Variable Region Amino Acid 10 AC 10 LightChain-CDR1(L1) Nucleotide 11 AC 10 Light Chain-CDR1(L1) Amino Acid 12 AC10 Light Chain-CDR2(L2) Nucleotide 13 AC 10 Light Chain-CDR2(L2) AminoAcid 14 AC 10 Light Chain-CDR3(L3) Nucleotide 15 AC 10 LightChain-CDR3(L3) Amino Acid 16 HeFi-1 Heavy Chain Variable RegionNucleotide 17 HeFi-1 Heavy Chain Variable Region Amino Acid 18 HeFi-1Heavy Chain-CDR1(H1) Nucleotide 19 HeFi-1 Heavy Chain-CDR1(H1) AminoAcid 20 HeFi-1 Heavy Chain-CDR2(H2) Nucleotide 21 HeFi-1 HeavyChain-CDR2(H2) Amino Acid 22 HeFi-1 Heavy Chain-CDR3(H3) Nucleotide 23HeFi-1 Heavy Chain-CDR3(H3) Amino Acid 24 HeFi-1 Light Chain VariableRegion Nucleotide 25 HeFi-1 Light Chain Variable Region Amino Acid 26HeFi-1 Light Chain-CDR1(L1) Nucleotide 27 HeFi-1 Light Chain-CDR1(L1)Amino Acid 28 HeFi-1 Light Chain-CDR2(L2) Nucleotide 29 HeFi-1 LightChain-CDR2(L2) Amino Acid 30 HeFi-1 Light Chain-CDR3(L3) Nucleotide 31HeFi-1 Light Chain-CDR3(L3) Amino Acid 32 Human gamma I constant regionAmino Acid 33 Human kappa constant region Amino Acid 34

In a specific embodiment, the invention encompasses an antibodycomprising a heavy chain variable domain, said variable domaincomprising (a) a set of three CDRs, in which said set of CDRs comprisesSEQ ID NO:4, 6, or 8 or comprises amino acid sequences that aresubstantially identical to the amino acid sequences set forth in SEQ IDNO:4, 6, or 8 and (b) a set of four framework regions, in which said setof framework regions differs from the set of framework regions in murinemonoclonal antibody AC:10, and in which said antibody immunospecificallybinds CD30.

In a specific embodiment, the invention encompasses an antibodycomprising a heavy chain variable domain, said variable domaincomprising (a) a set of three CDRs, in which said set of CDRs comprisesSEQ ID NO:20, 22 or 24 or comprises amino acid sequences that aresubstantially identical to the amino acid sequences set forth in SEQ IDNO:20, 22, or 24 and (b) a set of four framework regions, in which saidset of framework regions differs from the set of framework regions inmurine monoclonal antibody HeFi-1, and in which said antibodyimmunospecifically binds CD30.

In a specific embodiment, the invention encompasses an antibodycomprising a light chain variable domain, said variable domaincomprising (a) a set of three CDRs, in which said set of CDRs comprisesSEQ ID NO:12, 14 or 16 or comprises amino acid sequences that aresubstantially identical to the amino acid sequences set forth in SEQ IDNO:12, 14, or 16, and (b) a set of four framework regions, in which saidset of framework regions differs from the set of framework regions inmurine monoclonal antibody AC10, and in which said antibodyimmunospecifically binds CD30.

In a specific embodiment, the invention encompasses an antibodycomprising a light chain variable domain, said variable domaincomprising (a) a set of three CDRs, in which said set of CDRs comprisesSEQ ID NO:28, 30, or 32 or comprises amino acid sequences that aresubstantially identical to the amino acid sequences set forth in SEQ IDNO:28, 30, or 32, and (b) a set of four framework regions, in which saidset of framework regions differs from the set of framework regions inmurine monoclonal antibody HeFi-1, and in which said antibodyimmunospecifically binds CD30.

In certain embodiments, the anti-CD30 antibody comprises a gamma Iconstant region, (e.g., huCγl, SwissProt accession number P01857,incorporated herein by reference in its entirety) and a human kappaconstant region (e.g., huCκ, PID G185945, incorporated herein byreference in its entirety).

The present invention encompasses embodiments wherein a chimeric AC10antibody comprises the heavy chain variable region set forth in SEQ IDNO:2, the light chain variable region set forth in SEQ ID NO:10, thehuman gamma I constant region set forth in SEQ ID NO:33 (or amino acids1 to 329 of SEQ ID NO:33) and the human kappa constant region set forthin SEQ ID NO:34.

Additionally, the antibodies can also be described or specified in termsof their primary structures. In some embodiments, the variable regionsof the anti-CD30 antibodies will have at least 80%, at least 85%, atleast 90%, at least 95% and most preferably at least 98% identity (ascalculated using methods known in the art and described herein) to thevariable regions of murine AC10 or HeFi-1.

Exemplary anti-CD30 antibodies include functional derivatives or analogsof AC10 and HeFi-1. As used herein, the term “functional” in thiscontext indicates that the functional derivate or analog of AC10 andHeFi-1 is capable of specific binding to CD30.

Antibodies for use in the present invention include those thatcompetitively inhibit binding of murine AC10 or HeFi-1 to CD30 asdetermined by any method known in the art for determining competitivebinding. For example, the antibody can inhibit binding of AC10 or HcFi-1to CD30 by at least 50%, at least 60%, at least 70%, at least 80%, atleast 85%, at least 90%, or even at least 95%.

G. Antibody-Drug Conjugates

The methods described herein encompass the use of antibody drugconjugates comprising an anti-CD30 antibody, covalently linked to MMAEthrough a vc-PAB linker. The antibody drug conjugates are delivered tothe subject as a pharmaceutical composition.

The antibody-drug conjugates of the present invention have the followingformula:

or a pharmaceutically acceptable salt thereof;

wherein:

mAb is an anti-CD30 antibody,

S is a sulfur atom of the antibody

A- is a Stretcher unit,

p is from about 3 to about 5.

The drug loading is represented by p, the average number of drugmolecules per antibody in a pharmaceutical composition. For example, ifp is about 4, the average drug loading taking into account all of theantibody present in the pharmaceutical composition is about 4. P rangesfrom about 3 to about 5, more preferably from about 3.6 to about 4.4,even more preferably from about 3.8 to about 4.2. P can be about 3,about 4, or about 5. The average number of drugs per antibody inpreparation of conjugation reactions may be characterized byconventional means such as mass spectroscopy, ELISA assay, and HPLC. Thequantitative distribution of antibody-drug conjugates in terms of p mayalso be determined. In some instances, separation, purification, andcharacterization of homogeneous antibody-drug-conjugates where p is acertain value from antibody-drug-conjugates with other drug loadings maybe achieved by means such as reverse phase HPLC or electrophoresis.

The Stretcher unit (A), is capable of linking an antibody unit to thevaline-citrulline amino acid unit via a sulfhydryl group of theantibody. Sulfhydryl groups can be generated, for example, by reductionof the interchain disulfide bonds of an anti-CD30 antibody. For example,the Stretcher unit can be linked to the antibody via the sulfur atomsgenerated from reduction of the interchain disulfide bonds of theantibody. In some embodiments, the Stretcher units are linked to theantibody solely via the sulfur atoms generated from reduction of theinterchain disulfide bonds of the antibody. In some embodiments,sulfhydryl groups can be generated by reaction of an amino group of alysine moiety of an anti-CD30 antibody with 2-iminothiolane (Traut'sreagent) or other sulfhydryl generating reagents. In certainembodiments, the anti-CD30 antibody is a recombinant antibody and isengineered to carry one or more lysines. In certain other embodiments,the recombinant anti-CD30 antibody is engineered to carry additionalsulfhydryl groups, e.g., additional cysteines.

The synthesis and structure of MMAE is described in U.S. Pat. No.6,884,869 incorporated by reference herein in its entirety and for allpurposes. The synthesis and structure of exemplary Stretcher units andmethods for making antibody drug conjugates are described in, forexample, U.S. Publication Nos. 2006/0074008 and 2009/0010945 each ofwhich is incorporated herein by reference in its entirety.

Representative Stretcher units are depicted within the square bracketsof Formulas Ma and IIIb, wherein L-, —W—, —Y—, -D, w and y are asdefined above, and R₁₇ is selected from —C₁-C₁₀ alkylene-, —C₁-C₁₀alkenylene-, —C₁-C₁₀ alkynylene-, -carbocyclo-, —O—(C₁-C₈ alkylene)-,O—(C₁-C₈ alkenylene)-, —O—(C₁-C₈ alkynylene)-, -arylene-, —C₁-C₁₀alkylene-arylene-, —C₂-C₁₀ alkenylene-arylene, —C₂-C₁₀alkynylene-arylene, -arylene-C₁-C₁₀ alkylene-, -arylene-C₂-C₁₀alkenylene-, -arylene-C₂-C₁₀ alkynylene-, —C₁-C₁₀alkylene-(carbocyclo)-, —C₂-C₁₀ alkenylene-(carbocyclo)-, —C₂-C₁₀alkynylene-(carbocyclo)-, -(carbocyclo)-C₁-C₁₀ alkylene-,-(carbocyclo)-C₂-C₁₀ alkenylene-, -(carbocyclo)-C₂-C₁₀ alkynylene,-heterocyclo-, —C₁-C₁₀ alkylene-(heterocyclo)-, —C₂-C₁₀alkenylene-(heterocyclo)-, —C₂-C₁₀ alkynylene-(heterocyclo)-,-(heterocyclo)-C₁-C₁₀ alkylene-, -(heterocyclo)-C₂-C₁₀ alkenylene-,-(heterocyclo)-C₁-C₁₀ alkynylene-, —(CH₂CH₂O)_(r) or—(CH₂CH₂O)_(r)—CH₂—, and r is an integer ranging from 1-10, wherein saidalkyl, alkenyl, alkynyl, alkylene, alkenylene, alkynyklene, aryl,carbocyle, carbocyclo, heterocyclo, and arylene radicals, whether aloneor as part of another group, are optionally substituted.

An illustrative Stretcher unit is that of Formula Ma wherein R¹⁷ is—(CH₂)₅—:

Another illustrative Stretcher unit is that of Formula Ma wherein R¹⁷ is—(CH₂CH₂O)_(r)—CH₂—; and r is 2:

An illustrative Stretcher unit is that of Formula Ma wherein R¹⁷ is-arylene- or arylene-C₁-C₁₀ alkylene-. In some embodiments, the arylgroup is a phenyl group.

Still another illustrative Stretcher unit is that of Formula IIIbwherein R¹⁷ is —(CH₂)₅—:

One way that the Stretcher unit can be linked to the antibody unit isvia a disulfide bond between a sulfur atom of the antibody unit and asulfur atom of the Stretcher unit. In some embodiments, the sulfur atomis from an internal cysteine residue of the antibody. In some otherembodiments, the sulfur atom is from a cysteine residue that has beenengineered into the antibody.

A representative Stretcher unit is depicted within the square bracketsof Formula IV, wherein R¹⁷ is as defined above.

It should be noted that throughout this application, the S moiety in theformula below refers to a sulfur atom of the antibody, unless otherwiseindicated by context.

In a particularly preferred embodiment, of the present invention, thestrectcher is of Formula Ina and the antibody drug conjugate is aMC-vc-PAB-MMAE antibody drug conjugate as follows wherein p is fromabout 3 to about 5, preferably from 3.8 to 4.2. The S moiety refers to asulfur atom of the antibody.

In a particularly preferred embodiment, the antibody is a chimeric orhumanized AC10 antibody. In a particularly preferred embodiment, the Smoiety is generated by reduction of the interchain disulfide bonds ofthe antibody.

H. Pharmaceutical Compositions and Formulations

Various delivery systems can be used to administer antibody-drugconjugates. In certain preferred embodiments of the present invention,administration of the antibody-drug conjugate compound is by intravenousinfusion. In some embodiments, administration is by a two hourintravenous infusion.

The antibody-drug conjugate compound can be administered as apharmaceutical composition comprising one or more pharmaceuticallycompatible ingredients. For example, the pharmaceutical compositiontypically includes one or more pharmaceutically acceptable carriers, forexample, water-based carriers (e.g., sterile liquids). Water is a moretypical carrier when the pharmaceutical composition is administeredintravenously.

The composition, if desired, can also contain, for example, salinesalts, buffers, salts, nonionic detergents, and/or sugars. Examples ofsuitable pharmaceutical carriers are described in “Remington'sPharmaceutical Sciences” by E. W. Martin. The formulations correspond tothe mode of administration.

The present invention provides, for example, pharmaceutical compositionscomprising a therapeutically effective amount of the antibody-drugconjugate, a buffering agent, optionally a cryoprotectant, optionally abulking agent, optionally a salt, and optionally a surfactant.Additional agents can be added to the composition. A single agent canserve multiple functions. For example, a sugar, such as trehalose, canact as both a cryoprotectant and a bulking agent. Any suitablepharmaceutically acceptable buffering agents, surfactants,cyroprotectants and bulking agents can be used in accordance with thepresent invention.

In addition to providing methods for treating a CD30-expressing cancer,the present invention provides antibody drug conjugate formulationsincluding drug conjugate formulations that have undergonelyophilization, or other methods of protein preservation, as well asantibody drug formulations that have not undergone lyophilization.

In some embodiments, the antibody drug conjugate formulation comprises(i) about 1-25 mg/ml, preferably about 3 to about 10 mg/ml of anantibody-drug conjugate (e.g., an antibody-drug conjugate of formula Ior a pharmaceutically acceptable salt thereof), (ii) about 5-50 mM,preferably about 10 mM to about 25 mM of a buffer selected from acitrate, phosphate, or histidine buffer or combinations thereof,preferably sodium citrate, potassium phosphate, histidine, histidinehydrochloride, or combinations thereof, (iii) about 3% to about 10%sucrose or trehalose or combinations thereof, (iv) optionally about 0.05to 2 mg/ml of a surfactant selected from polysorbate 20 or polysorbate80 or combinations thereof; and (v) water, wherein the pH of thecomposition is from about 5.3 to about 7, preferably about 6.6.

In some embodiments, an antibody drug conjugate formulation willcomprise about 1-25 mg/ml, preferably about 3 to about 10 mg/ml, evenmore preferably about 5 mg/ml of an antibody-drug conjugate, (ii) about10 mM to about 25 mM of a buffer selected from sodium citrate, potassiumphosphate, histidine, histidine hydrochloride or combinations thereof,(iii) about 3% to about 7% trehalose or sucrose or combinations thereof,optionally (iv) about 0.05 to about 1 mg/ml of a surfactant selectedfrom polysorbate 20 or polysorbate 80, and (v) water, wherein the pH ofthe composition is from about 5.3 to about 7, preferably about 6.6.

In some embodiments, an antibody drug conjugate formulation willcomprise about 5 mg/ml of an antibody-drug conjugate, (ii) about 10 mMto about 25 mM of a buffer selected from sodium citrate, potassiumphosphate, histidine, histidine hydrochloride or combinations thereof,(iii) about 3% to about 7% trehalose, optionally (iv) about 0.05 toabout 1 mg/ml of a surfactant selected from polysorbate 20 orpolysorbate 80, and (v) water, wherein the pH of the composition is fromabout 5.3 to about 7, preferably about 6.6.

The antibody drug conjugate formulation can, for example, comprise (orconsist essentially of) about 5 mg/ml of an antibody-drug conjugate,(ii) about 20 mM sodium citrate (iii) about 6% to about 7% trehalose(about 70 mg/ml), (iv) about 0.1 to 0.3 mg/ml of a surfactant selectedfrom polysorbate 20 or polysorbate 80 (preferably polysorbate 80), and(v) water, wherein the pH of the composition is from about 5.3 to about7, preferably about 6.6.

Any of the formulations described above can be stored in a liquid orfrozen form and can be optionally subjected to a preservation process.In some embodiments, the formulations described above are lyophilized,i.e., they are subjected to lyophilization. In some embodiments, theformulations described above are subjected to a preservation process,for example, lyophilization, and are subsequently reconstituted with asuitable liquid, for example, water. By lyophilized it is meant that thecomposition has been freeze-dried under a vacuum. Lyophilizationtypically is accomplished by freezing a particular formulation such thatthe solutes are separated from the solvent(s). The solvent is thenremoved by sublimation (i.e., primary drying) and next by desorption(i.e., secondary drying).

In some embodiments, lyophilized formulations of the present inventioncomprise the antibody-drug conjugate, a buffering agent, optionally acryoprotectant, optionally a bulking agent, optionally a salt, andoptionally a surfactant as well as additional agents, wherein thecomposition has a pH of about 5.3-7 when reconstituted with water. Insome embodiments, lyophilized formulations of the present invention,when reconstituted with water, comprise (i) about 1-25 mg/ml, preferablyabout 3 to about 10 mg/ml, of an antibody-drug conjugate (e.g., anantibody-drug conjugate of formula I or a pharmaceutically acceptablesalt thereof), (ii) about 5-50 mM, preferably about 10 mM to about 25 mMof a buffer selected from a citrate, phosphate, or histidine buffer orcombinations thereof, preferably sodium citrate, potassium phosphate,histidine, histidine hydrochloride, or combinations thereof, (iii) about3% to about 10% sucrose or trehalose or combinations thereof, and (iv)optionally about 0.05 to 2 mg/ml of a surfactant selected frompolysorbate 20 or polysorbate 80 or combinations thereof; wherein the pHof the composition is from about 5.3 to about 7, preferably about 6.6.

In some embodiments, lyophilized formulations, when reconstituted withwater, comprise (i) about 1-25 mg/ml, preferably about 3 to about 10mg/ml, even more preferably about 5 mg/ml of an antibody-drug conjugate(ii) about 10 mM to about 25 mM of a buffer selected from sodiumcitrate, potassium phosphate, histidine, histidine hydrochloride orcombinations thereof, (iii) about 3% to about 7% trehalose or sucrose orcombinations thereof, and optionally (iv) about 0.05 to about 1 mg/ml ofa surfactant selected from polysorbate 20 or polysorbate 80, wherein thepH of the composition is from about 5.3 to about 7, preferably about6.6.

In some embodiments, lyophilized formulations, when reconstituted withwater, comprise about 5 mg/ml of an antibody-drug conjugate of thepresent invention, (ii) about 10 mM to about 25 mM of a buffer selectedfrom sodium citrate, potassium phosphate, histidine, histidinehydrochloride, or combinations thereof, (iii) about 3% to about 7%trehalose, optionally (iv) about 0.05 to about 1 mg/ml of a surfactantselected from polysorbate 20 or polysorbate 80, wherein the pH of thecomposition is from about 5.3 to about 7, preferably about 6.6.

In some embodiments, lyophilized formulations, when reconstituted withwater, comprise (or consist essentially of) about 5 mg/ml of anantibody-drug conjugate of the present invention, (ii) about 20 mMsodium citrate (iii) about 6% to about 7% trehalose (about 70 mg/ml),(iv) about 0.1 to 0.3 mg/ml of a surfactant selected from polysorbate 20or polysorbate 80 (preferably polysorbate 80), wherein the pH of thecomposition is from about 5.3 to about 7, preferably about 6.6.

The formulations of the present invention can be used with the methodsdescribed herein or with other methods for treating disease. Theantibody drug conjugate formulations may be further diluted beforeadministration to a subject. In some embodiments, the formulations willbe diluted with saline and held in IV bags or syringes beforeadministration to a subject. Accordingly, in some embodiments, themethods for treating a CD30-expressing hematologic cancer in a subjectwill comprise administering to a subject in need thereof a weekly doseof a pharmaceutical composition comprising antibody-drug conjugateshaving formula I wherein the administered dose of antibody-drugconjugates is from about 0.8 mg/kg of the subject's body weight to about1.8 mg/kg of the subject's body weight and the pharmaceuticalcomposition is administered for at least three weeks and wherein theantibody drug conjugates, prior to administration to a subject, werepresent in a formulation comprising (i) about 1-25 mg/ml, preferablyabout 3 to about 10 mg/ml of the antibody-drug conjugate (ii) about 5-50mM, preferably about 10 mM to about 25 mM of a buffer selected fromsodium citrate, potassium phosphate, histidine, histidine hydrochloride,or combinations thereof, (iii) about 3% to about 10% sucrose ortrehalose or combinations thereof, (iv) optionally about 0.05 to 2 mg/mlof a surfactant selected from polysorbate 20 or polysorbate 80 orcombinations thereof; and (v) water, wherein the pH of the compositionis from about 5.3 to about 7, preferably about 6.6.

In typical embodiments, the pharmaceutical composition is formulated inaccordance with routine procedures as a pharmaceutical compositionadapted for intravenous administration to human beings. Typically,compositions for intravenous administration are solutions in sterileisotonic aqueous buffer. Where necessary, the pharmaceutical can alsoinclude a solubilizing agent and a local anesthetic such as lignocaineto ease pain at the site of the injection. Generally, the ingredientsare supplied either separately or mixed together in unit dosage form,for example, as a dry lyophilized powder or water free concentrate in ahermetically sealed container such as an ampoule or sachette indicatingthe quantity of active agent. Where the pharmaceutical is to beadministered by infusion, it can be dispensed, for example, with aninfusion bottle containing sterile pharmaceutical grade water or saline.Where the pharmaceutical is administered by injection, an ampoule ofsterile water for injection or saline can be, for example, provided sothat the ingredients can be mixed prior to administration.

The present invention also provides kits for the treatment of aCD30-expressing hematologic cancer. The kit can comprise (a) a containercontaining the antibody-drug conjugate. Such kits can further include,if desired, one or more of various conventional pharmaceutical kitcomponents, such as, for example, containers with one or morepharmaceutically acceptable carriers, additional containers, etc., aswill be readily apparent to those skilled in the art. Printedinstructions, either as inserts or as labels, indicating quantities ofthe components to be administered, guidelines for administration, and/orguidelines for mixing the components, can also be included in the kit.

The present invention is not to be limited in scope by the specificembodiments described herein. Various modifications of the invention inaddition to those described herein will become apparent to those skilledin the art from the foregoing description and accompanying figures. Suchmodifications are intended to fall within the scope of the appendedclaims.

All publications and patent documents cited above are herebyincorporated by reference in their entirety for all purposes to the sameextent as if each were so individually denoted.

The invention is further described in the following examples, which arein not intended to limit the scope of the invention.

EXAMPLES

A multicenter, phase 1, weekly dosing, dose escalation study wasconducted in pts with refractory or relapsed HL or systemic ALCL. ThecAC10-MC-vc-PAB-MMAE antibody drug conjugate was administered weekly atdoses of 0.4-1.4 mg/kg (2-hr IV infusions). Patients with standarddisease or better after two 21-day cycles (6 doses) were eligible tocontinue treatment with the antibody drug conjugate. The weekly studydesign is provided in FIG. 1.

Results: In 34 patients, median age was 34 yrs (range 13-82). Patientsreceived a median of 5 prior therapies; 62% received an autologous SCT.MTD was exceeded at 1.4 mg/kg. Exposure to SGN-35 (AUC) increasedrelative to dose level. Multiple CRs were observed at higher doses(table); observed time to response for 1 mg/kg dose was approximately 8wks.

TABLE 2 Best Clinical Response* Dose Group (mg/kg) N CR PR SD PD 0.4 4 ——  4⁺ — 0.6 3 — 1 1 1 0.8 6 4⁺⁺ —  1^(P) 1 1.0 6 4⁺⁺ 1 1 — 1.2 5 — 1 3^(P) 1 1.4 3  2 — 1 — Total 27 10 3 22  3 *Based on InternationalWorking Group Revised Response Criteria for Malignant Lymphoma (Cheson,2007). 7 patients not evaluable (pending Cycle 2 restage). +represents 1patient with ALCL; ++represents 2 patient with ALCL; each P represents 1pediatric patient (12-17 years)

TABLE 3 Percent Response Response Rate All Patients HL Patients sALCLpatients Overall Response 48% (13/27) 41% (9/22) 80% (4/5) Rate (CR +PR) Complete Response 37% (10/27) 27% (6/22) 80% (4/5) Tumor Reductions81% (22/27)

The cAC10-MC-vc-PAB-MMAE antibody drug conjugate was generally welltolerated in the 34 treated patients. There were no grade 5 events. Thegrade 4 events were neutropenia (1 subject), and hyperglycemia (1). Thegrade 3 events were neutropenia (3), diarrhea (1), paresthesia (1),vomiting (1), and leucopenia (1). The grade 2 or lower adverse eventswere nausea (9), fatigue (8), peripheral neuropathy (6), neutropenia(2), dizziness (4), hyperglycemia (3), and paresthesia (3). The doselimiting toxicity was at 1.4 mg/kg.

SEQUENCE LISTING SEQ ID NO: 1cag atc cag ctg cag cag tct gga cct gag gtg gtg aag cct ggg gcttca gtg aag ata tcc tgc aag gct tct ggc tac acc ttc act gac tactat ata acc tgg gtg aag cag aag cct gga cag gga ctt gag tgg attgga tgg att tat cct gga agc ggt aat act aag tac aat gag aag ttcaag ggc aag gcc aca ttg act gta gac aca tcc tcc agc aca gcc ttcatg cag ctc agc agc ctg aca tct gag gac act gct gtc tat ttc tgtgcg aac tat ggt aac tac tgg ttt gct tac tgg ggc caa ggg act caggtc act gtc tct gca SEQ ID NO: 2Gln Ile Gln Leu Gln Gln Ser Gly Pro Glu Val Val Lys Pro Gly AlaSer Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp TyrTyr Ile Thr Trp Val Lys Gln Lys Pro Gly Gln Gly Leu Glu Trp IleGly Trp Ile Tyr Pro Gly Ser Gly Asn Thr Lys Tyr Asn Glu Lys PheLys Gly Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser Thr Ala PheMet Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Phe CysAla Asn Tyr Gly Asn Tyr Trp Phe Ala Tyr Trp Gly Gln Gly Thr GlnVal Thr Val Ser Ala SEQ ID NO: 3 gactactata taacc SEQ ID NO: 4Asp Tyr Tyr Ile Thr SEQ ID NO: 5tggatttatc ctggaagcgg taatactaag tacaatgaga agttcaaggg c SEQ ID NO: 6Trp Ile Tyr Pro Gly Ser Gly Asn Thr Lys Tyr Asn Glu Lys Phe Lys GlySEQ ID NO: 7 tatggtaact actggtttgc ttac 24 SEQ ID NO: 8Tyr Gly Asn Tyr Trp Phe Ala Tyr SEQ ID NO: 9gac att gtg ctg acc caa tct cca gct tct ttg gct gtg tct cta gggcag agg gcc acc atc tcc tgc aag gcc agc caa agt gtt gat ttt gatggt gat agt tat atg aac tgg tac caa cag aaa cca gga cag cca cccaaa gtc ctc atc tat gct gca tcc aat cta gaa tct ggg atc cca gccagg ttt agt ggc agt ggg tct ggg aca gac ttc acc ctc aac atc catcct gtg gag gag gag gat gct gca acc tat tac tgt cag caa agt aatgag gat ccg tgg acg ttc ggt gga ggc acc aag ctg gaa atc aaaSEQ ID NO: 10Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu GlyGln Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Phe AspGly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Pro ProLys Val Leu Ile Tyr Ala Ala Ser Asn Leu Glu Ser Gly Ile Pro AlaArg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile HisPro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Ser AsnGlu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile LysSEQ ID NO: 11 aaggccagcc aaagtgttga ttttgatggt gatagttata tgaacSEQ ID NO: 12Lys Ala Ser Gln Ser Val Asp Phe Asp Gly Asp Ser Tyr Met AsnSEQ ID NO: 13 gctgcatcca atctagaatc t SEQ ID NO: 14Ala Ala Ser Asn Leu Glu Ser SEQ ID NO: 15 cagcaaagta atgaggatcc gtggacgSEQ ID NO: 16 Gln Gln Ser Asn Glu Asp Pro Trp Thr SEQ ID NO: 17gag gtg aag ctg gtg gag tct gga gga ggc ttg gta cag cct ggg ggttct ctg aga ctc tcc tgt gca act tct ggg ttc acc ttc agt gat tactat atg aac tgg gtc cgc cag cct cca gga aag gct ctt gag tgg ttgggt ttt att aga aac aaa gct aat ggt tac aca aca gag ttc agt gcatct gtg atg ggt cgg ttc acc atc tcc aga gat gat tcc caa agc atcctc tat ctt cag atg aac acc ctg aga gct gag gac agt gcc act tattac tgt gca aga gat ccc ccc tat ggt aac ccc cat tat tat gct atggac tac tgg ggt caa gga acc tca gtc acc gtc tcc tca SEQ ID NO: 18Glu Val Lys Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly GlySer Leu Arg Leu Ser Cys Ala Thr Ser Gly Phe Thr Phe Ser Asp TyrTyr Met Asn Trp Val Arg Gln Pro Pro Gly Lys Ala Leu Glu Trp LeuGly Phe Ile Arg Asn Lys Ala Asn Gly Tyr Thr Thr Glu Phe Ser AlaSer Val Met Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Gln Ser IleLeu Tyr Leu Gln Met Asn Thr Leu Arg Ala Glu Asp Ser Ala Thr TyrTyr Cys Ala Arg Asp Pro Pro Tyr Gly Asn Pro His Tyr Tyr Ala MetAsp Tyr Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser SEQ ID NO: 19gattactata tgaac SEQ ID NO: 20 Asp Tyr Tyr Met Asn SEQ ID NO: 21tttattagaa acaaagctaa tggttacaca acagagttca gtgcatctgt gatgggtSEQ ID NO: 22Phe Ile Arg Asn Lys Ala Asn Gly Tyr Thr Thr Glu Phe Ser Ala SerVal Met Gly SEQ ID NO: 23 gatcceccct atggtaaccc ccattattat gctatggact acSEQ ID NO: 24 Asp Pro Pro Tyr Gly Asn Pro His Tyr Tyr Ala Met Asp TyrSEQ ID NO: 25gac att gtg ctg acc cag tct cct gct tcc tta gct gtt tct ctg gggcag agg gcc acc atc tca tgc agg gcc agc aaa agt gtc agt gca tctggc tat aat tat atg cac tgg tac caa cag aaa gca ggg cag cca cccaaa ctc ctc atc cat ctt gca tcc aac cta gaa tct ggg gtc cct gccagg ttc agt ggc agt ggg tct ggg aca gac ttc acc ctc aac atc catcct gtg gag gag gag gat gct tca acc tat tac tgt cag cac agt ggggag ctt cca ttc acg ttc ggc tcg ggg aca aag ttg gaa ata aaaSEQ ID NO: 26Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu GlyGln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys Ser Val Ser Ala SerGly Tyr Asn Tyr Met His Trp Tyr Gln Gln Lys Ala Gly Gln Pro ProLys Leu Leu Ile His Leu Ala Ser Asn Leu Glu Ser Gly Val Pro AlaArg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile HisPro Val Glu Glu Glu Asp Ala Ser Thr Tyr Tyr Cys Gln His Ser GlyGlu Leu Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile LysSEQ ID NO: 27 agggccagca aaagtgtcag tgcatctggc tataattata tgcacSEQ ID NO: 28Arg Ala Ser Lys Ser Val Ser Ala Ser Gly Tyr Asn Tyr Met HisSEQ ID NO: 29 cttgcatcca acctagaatc t SEQ ID NO: 30Leu Ala Ser Asn Leu Glu Ser SEQ ID NO: 31 cageacagtg gggagettec atteacgSEQ ID NO: 32 Gln His Ser Gly Glu Leu Pro Phe Thr SEQ ID NO: 33Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser LysSer Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp TyrPhe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr SerGly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr SerLeu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln ThrTyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp LysLys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro CysPro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro ProLys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr CysVal Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn TrpTyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg GluGlu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Lou Thr Val LouHis Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser AsnLys Ala Lou Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys GlyGln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp GluLeu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe TyrPro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu AsnAsn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe PheLeu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly AsnVal Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr ThrGln Lys Ser Leu Ser Leu Ser Pro Gly Lys SEQ ID NO: 34Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu GlnLeu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe TyrPro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln SerGly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser ThrTyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu LysHis Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser ProVal Thr Lys Ser Phe Asn Arg Gly Glu Cys

1-25. (canceled)
 26. A method for treating a CD30-expressing hematologiccancer in a human subject, the method comprising administering to asubject in need thereof a dose of a pharmaceutical compositioncomprising cAC10-MC-vc-PAB-MMAE antibody-drug conjugate, wherein theadministered dose of cAC10-MC-vc-PAB-MMAE antibody-drug conjugate is 1.2mg/kg of the subject's body weight and the pharmaceutical composition isadministered once every two weeks, and wherein the CD30-expressinghematologic cancer is Hodgkin Lymphoma.
 27. The method of claim 26,wherein the human subject is newly diagnosed and has not previously beentreated for the CD30-expressing hematologic cancer.
 28. The method ofclaim 26, wherein the pharmaceutical composition is administered for twoor more treatment cycles.
 29. The method of claim 28, wherein thepharmaceutical composition is administered for 3 or more treatmentcycles.
 30. The method of claim 28, wherein the pharmaceuticalcomposition is administered for 4 or more treatment cycles.
 31. Themethod of claim 28, wherein the pharmaceutical composition isadministered for 5 or more treatment cycles.
 32. A method for treating aCD30-expressing hematologic cancer in a human subject, the methodcomprising administering to a subject in need thereof a dose of apharmaceutical composition comprising cAC10-MC-vc-PAB-MMAE antibody-drugconjugate, wherein the administered dose of cAC10-MC-vc-PAB-MMAEantibody-drug conjugate is 1.2 mg/kg of the subject's body weight andthe pharmaceutical composition is administered once every two weeks, andwherein the average number of MMAE molecules per cAC10 antibody of thecAC10-MC-vc-PAB-MMAE antibody-drug conjugate in the pharmaceuticalcomposition is about
 4. 33. The method of claim 32, wherein the humansubject is newly diagnosed and has not previously been treated for theCD30-expressing hematologic cancer.
 34. The method of claim 32, whereinthe pharmaceutical composition is administered for two or more treatmentcycles.
 35. The method of claim 32, wherein the pharmaceuticalcomposition is administered for 3 or more treatment cycles.
 36. Themethod of claim 32, wherein the pharmaceutical composition isadministered for 4 or more treatment cycles.
 37. The method of claim 32,wherein the pharmaceutical composition is administered for 5 or moretreatment cycles.
 38. A method for treating a CD30-expressinghematologic cancer in a human subject, the method comprisingadministering to a subject in need thereof a dose of a pharmaceuticalcomposition comprising (i) cAC10-MC-vc-PAB-MMAE antibody-drug conjugate,wherein the administered dose of cAC10-MC-vc-PAB-MMAE antibody-drugconjugate is 1.2 mg/kg of the subject's body weight and thepharmaceutical composition is administered once every two weeks, whereinthe CD30-expressing hematologic cancer is Hodgkin Lymphoma, and whereinthe average number of MMAE molecules per cAC10 antibody of thecAC10-MC-vc-PAB-MMAE antibody-drug conjugate in the pharmaceuticalcomposition is about
 4. 39. The method of claim 38, wherein the humansubject is newly diagnosed and has not previously been treated for theCD30-expressing hematologic cancer.
 40. The method of claim 38, whereinthe pharmaceutical composition is administered for two or more treatmentcycles.
 41. The method of claim 38, wherein the pharmaceuticalcomposition is administered for 3 or more treatment cycles.
 42. Themethod of claim 38, wherein the pharmaceutical composition isadministered for 4 or more treatment cycles.
 43. The method of claim 38,wherein the pharmaceutical composition is administered for 5 or moretreatment cycles.